蛋鸡感染NDV后输卵管组织中CaBP-D28k、OVA基因mRNA表达及蛋品质量变化研究

发布时间：2018-05-31 01:04

本文选题：新城疫病毒 + 产蛋鸡　；参考：《西北农林科技大学》2015年硕士论文

【摘要】：新城疫是由新城疫病毒(NDV)引起禽类的一种急性、高度传染性疾病。NDV感染产蛋鸡群,除发生发病死亡外,还可引起生殖系统病变,导致鸡群产蛋量和蛋品质下降,产沙壳蛋、软壳蛋和畸形蛋等。但目前有关NDV感染蛋鸡后导致输卵管组织病变与蛋品质下降的机制还不清楚。本研究通过滴鼻、点眼途径对240日龄非免疫健康罗曼白蛋鸡感染NDV/Chicken/TC/1/2011株,在健康对照鸡和感染后第1、3、5、7、9、11、15天,收集健康对照鸡和感染鸡所产鸡蛋检测蛋壳质量,采用原子分光光度法检测蛋壳中钙含量,采用扫描电子显微镜观察鸡蛋蛋壳内、外表面和横断面超微结构;在健康对照组和感染后第1、3、5、7、9、11、15天,分别扑杀一定数量鸡,无菌采集输卵管膨大部、子宫部,利用实时荧光定量PCR检测与蛋壳钙化相关的CaBP-D28k基因和与蛋清质量相关的卵清蛋白(OVA)基因mRNA的表达变化,结果如下:1.蛋鸡感染NDV后1、3、5、7、9、11、15天,感染鸡所产鸡蛋蛋壳中钙的平均相对含量(质量比)分别40.60%、38.41%、34.15%、37.42%、38.58%、39.03%、39.50%,健康对照鸡为41.88%;感染鸡所产鸡蛋蛋壳横断面平均厚度分别305um、290um、279.5um、313um、320.50um、321um、323um,健康对照鸡为368.5um。感染NDV后1~5天,鸡蛋乳突层变薄,栅栏层和垂直晶体层的平整性和致密性均不同程度地下降;蛋壳外表面的角质层逐渐变粗糙,气孔增大,皲裂明显;蛋壳内表面纤维间隙逐渐增大,结构稀疏、断裂明显、其上的结点明显变少。感染NDV后7~15天,蛋鸡所产鸡蛋蛋壳质量缓慢改善。2.蛋鸡感染NDV后1~5天,感染鸡产软壳蛋、薄壳蛋,输卵管子宫部CaBPD28k基因mRNA表达量持续下降,感染第5天时其表达量仅为健康对照鸡的0.16倍;感染后7~15天,发病鸡产蛋蛋壳质量逐渐改善,子宫部CaBP-D28k基因mRNA表达量逐渐回升。感染后第15天,子宫部CaBP-D28k基因mRNA表达量达到健康对照鸡的0.45倍。3.蛋鸡感染NDV后1、3、5、7、9、11、15天,感染鸡所产鸡蛋蛋清中蛋白相对含量分别为10.39%、9.71%、9.32%、9.51%、9.75%、9.93%、10.35%,健康对照鸡为11.05%;蛋鸡感染NDV后1~5天,输卵管组织中膨大部OVA基因mRNA表达量持续下降,感染第5天其表达量仅为健康对照鸡的0.08倍;感染后7~15天,感染鸡输卵管膨大部OVA基因mRNA表达量逐渐回升,感染后第15天,OVA基因mRNA的表达量达到健康对照鸡的0.27倍。研究表明,NDV感染蛋鸡后,感染鸡所产鸡蛋蛋壳钙含量的变化与蛋壳质量、鸡输卵管子宫部CaBP-D28k基因mRNA的表达量呈正相关;鸡蛋蛋清蛋白含量变化与鸡输卵管膨大部OVA基因mRNA表达量也呈正相关。
[Abstract]:Newcastle disease (NDV) is an acute and highly infectious disease of poultry caused by Newcastle disease virus (NDV). Soft shell eggs and deformed eggs, etc. However, the mechanism of fallopian tube pathological changes and egg quality decline after NDV infection in laying hens is unclear. In this study, NDV/Chicken/TC/1/2011 strain was infected with 240-day-old non-immune healthy Roman-white laying hens by nasal drip and eye drops. The egg shell quality of healthy control and infected chickens was detected by collecting eggs produced by healthy control chickens and infected chickens on day 1, 3, 5, 7, 7, 1, 1115 days after infection. The content of calcium in egg shell was detected by atomic spectrophotometry, the ultrastructure of inside, outside and cross section of egg shell was observed by scanning electron microscope, and a certain number of chickens were culled in the healthy control group and the first day after infection. The changes of mRNA expression of CaBP-D28k gene associated with eggshell calcification and egg white quality related ovalbumin (OVA) gene were detected by real-time fluorescence quantitative PCR. The results were as follows: 1. The average relative content (mass ratio) of calcium in eggs produced by infected hens was 40.60,38.41mg / kg, 38.41mg / kg and 38.58.15mg / kg, respectively. The average thickness of eggs produced by infected hens was 35.80um279.5um279.5um313um320.50um321um323umrespectively, and that of healthy chickens was 38.58um39.03um39.50um. the average thickness of egg shells produced by infected chickens was 305um.290um279.5um.The average thickness of eggs produced by infected chickens was 313um320.50um321um323um. the average thickness of eggs produced by infected chickens was 313um320.50um321um323um. The papillomastoid layer thinned, the flatness and compactness of palisade layer and vertical crystal layer decreased in varying degrees, the cuticle on the outer surface of eggshell became rough, stomata enlarged and chapped obviously. The fiber gap on the inner surface of the eggshell gradually increases, the structure is sparse, the fracture is obvious, and the nodes on the eggshell are obviously reduced. The egg shell quality of laying hens improved slowly from 7 to 15 days after infection with NDV. The mRNA expression of CaBPD28k gene in the oviduct and uterus of the laying hens was decreased continuously on the 1st and 5th day after infection with NDV, and the mRNA expression of CaBPD28k gene was only 0.16 times of that of the healthy chickens on the 5th day after infection, and the expression level of CaBPD28k gene was only 0.16 times that of the healthy chickens on the 5th day after infection, and the expression level was 715 days after infection. The quality of egg shell was improved and the expression of CaBP-D28k gene mRNA in uterus was gradually increased. On the 15th day after infection, the expression of CaBP-D28k mRNA in uterus was 0.45 times as high as that in healthy control. The relative contents of protein in egg white of infected hens were 10.39 ~ 9.71and 9.32 ~ 9.75 ~ 9.75 ~ 10.35 days after infection with NDV and 11.05 ~ 15 days after NDV infection in laying hens, the expression of OVA gene mRNA in oviduct tissues of laying hens continued to decrease, and the expression of OVA gene mRNA in oviducts of oviduct tissues continued to decrease, while that of healthy control chickens was 11.05%, and the expression of OVA gene in the oviducts of oviducts continued to decrease after 15 days of infection, while that of healthy control chickens was 11.05%. On the 5th day of infection, the expression level of OVA gene was only 0.08 times of that of the healthy control, and the expression of OVA gene mRNA increased gradually on the 7th and 15th day after infection, and on the 15th day after infection, the expression of mRNA was 0.27 times higher than that of the healthy control. The results showed that there was a positive correlation between the changes of egg shell calcium content and egg shell quality and the expression of CaBP-D28k gene mRNA in oviduct uterus. The change of egg white protein content was also positively correlated with the expression of OVA gene mRNA in the enlarged oviduct.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.31

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